| Project/Area Number |
14035222
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| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas
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| Allocation Type | Single-year Grants |
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| Review Section |
Biological Sciences
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| Research Institution | Niigata University (2003-2006)
Shinshu University (2002) |
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Principal Investigator |
UCHIUMI Toshio Niigata University, Institute of Science and Technology, Professor (50143764)
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| Project Period (FY) |
2002 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥62,600,000 (Direct Cost: ¥62,600,000)
Fiscal Year 2006: ¥12,400,000 (Direct Cost: ¥12,400,000)
Fiscal Year 2005: ¥12,400,000 (Direct Cost: ¥12,400,000)
Fiscal Year 2004: ¥12,600,000 (Direct Cost: ¥12,600,000)
Fiscal Year 2003: ¥12,600,000 (Direct Cost: ¥12,600,000)
Fiscal Year 2002: ¥12,600,000 (Direct Cost: ¥12,600,000)
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| Keywords | Ribosome / GTPase / Ribosomal protein / rRNA / Translation / Elongation factor / RNA-protein interaction / Archaebacteria / RNA-タンバク質相互作用 / GTPase / IRES / 翻訳 / 蚕 / リボソームRNA / リボソームPタンパク質 / L7 / L12 / L10 |
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| Research Abstract |
The ribosomal GTPase-associated center is composed of a part of 23S rRNA and a few proteins, which assemble onto the rRNA region and form a characteristic complex, termed the stalk. This rRNA-protein complex plays a crucial role in ribosomal interaction with GTP binding translation factors, GTP hydrolysis, and regulation of translation rate. In this project, I analyzed the complex structure by biochemical approaches, and obtained the following new information, particularly on the stalk complex.
1) Establishment of a functional assay system, "hybrid ribosome"
We established conditions for in vitro reconstitution of the stalk complex in Escherichia coli ribosomes and its replacement with its counterparts from other species. This provided a useful system to investigate functional structure of the stalk complex.
2) Assembly mode of eukaryotic PO. P1-P2 stalk complex and its functional characterization
We demonstrated that eukaryotic proteins P1 and P2 form a heterodimer and two dimes bind to the C-terminal region of P0 to form pentameric complex. We also clarified that the stalk complex modulates functional structure of 23S rRNA in the GTPase-associated center.
3) Structural and functional characterization of the archaebacterial stalk complex
We demonstrated that archaebacterial L12 protein forms a homodimer and three dimers bind to the C-terminal region of the anchor protein P0, and that the resultant heptameric complex shows accessibility to eukaryotic as well as archaebacterial translation factors.
4) Structural and functional characterization of the eubacterial stalk complex
We showed that two E. coli L12 homodimers bind to the C-terminal region of the anchor protein L10, whereas three L12 dimers bind to L10 in case of thermophilic eubacteria. We also clarified that a complex variant composed of L10 and one L12 dimer is unstable on the ribosome. This suggests a relationship between number of L12 dimer and stability of the stalk complex in the ribosome.
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